三七总皂苷对大鼠胚胎皮质神经干细胞增殖和分化的影响。

Effects of Panax notoginseng saponins on proliferation and differentiation of rat embryonic cortical neural stem cells.

作者信息

Si Yinchu, Zhu Jun, Huang Xiang, Zhu Peichun, Xie Chune

机构信息

Department of Anatomy, Beijing University of Chinese Medicine, Beijing, China.

Department of Laboratory Medicine, University of California, San Francisco, CA, USA.

出版信息

J Chin Med Assoc. 2016 May;79(5):256-63. doi: 10.1016/j.jcma.2015.10.011. Epub 2016 Feb 22.

DOI:10.1016/j.jcma.2015.10.011

PMID:26915440

Abstract

BACKGROUND

We aimed to study the effect of Panax notoginseng saponins (PNS) on the proliferation, differentiation, self-renewal, and expressions of basic fibroblast growth factor (bFGF) and brain-derived neurotrophic factor (BDNF) in rat embryonic neural stem cells (NSCs).

METHODS

Cortical stem cells were isolated from rat embryos on Embryonic Day 17 (E17) and identified by nestin expression. Subsequently, primary culture, subculturing, and single cell cloning were performed on the cells. After the first cell passage (P1), the cells were resuspended and divided into a control group and a treatment group. Control cells were cultured in serum-free basal culture medium with B27 and dulbecco's modified eagle medium (DMEM)/F12. The same medium supplemented with PNS (100 μg/mL) was used to culture cells in the treatment group. Both groups were incubated at 37°C in a 5% CO2 incubator. Immunocytochemistry was performed 4 days after incubation.

RESULTS

Primary, P1, and P2 cells in the treatment group formed neurospheres, as did single cell clones of the P1 cells in this group. After being cultured for 4 days, the number of nestin-, proliferating cell nuclear antigen (PCNA)-, Tuj-1-, neurofilament (NF)-, vimentin-, glial fibrillary acidic protein (GFAP)-, bFGF-, and BDNF-positive cells significantly increased in the treatment group in comparison to the control group. All positively stained cells could form clear clusters.

CONCLUSION

PNS can promote rat embryonic cortical NSC survival, self-renewal, proliferation, and differentiation through neurotrophic factors by autocrine or paracrine signaling.

摘要

背景

我们旨在研究三七总皂苷（PNS）对大鼠胚胎神经干细胞（NSCs）增殖、分化、自我更新以及碱性成纤维细胞生长因子（bFGF）和脑源性神经营养因子（BDNF）表达的影响。

方法

在胚胎第17天（E17）从大鼠胚胎中分离皮质干细胞，并通过巢蛋白表达进行鉴定。随后，对这些细胞进行原代培养、传代培养和单细胞克隆。在首次细胞传代（P1）后，将细胞重悬并分为对照组和治疗组。对照细胞在含有B27和杜氏改良 Eagle培养基（DMEM）/F12的无血清基础培养基中培养。治疗组细胞在相同培养基中添加PNS（100μg/mL）进行培养。两组均在37°C、5%二氧化碳培养箱中孵育。孵育4天后进行免疫细胞化学检测。

结果

治疗组的原代、P1和P2细胞形成了神经球，该组P1细胞的单细胞克隆也形成了神经球。培养4天后，与对照组相比，治疗组中巢蛋白、增殖细胞核抗原（PCNA）、Tuj-1、神经丝（NF）、波形蛋白、胶质纤维酸性蛋白（GFAP）、bFGF和BDNF阳性细胞的数量显著增加。所有阳性染色细胞均可形成清晰的细胞簇。